CN100504378C - Method for preparing functional target disk of mass spectrum sample desorption by ground substance assistant laser - Google Patents
Method for preparing functional target disk of mass spectrum sample desorption by ground substance assistant laser Download PDFInfo
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- CN100504378C CN100504378C CNB2007100367896A CN200710036789A CN100504378C CN 100504378 C CN100504378 C CN 100504378C CN B2007100367896 A CNB2007100367896 A CN B2007100367896A CN 200710036789 A CN200710036789 A CN 200710036789A CN 100504378 C CN100504378 C CN 100504378C
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Abstract
A method for preparing mass spectrum sample function target plate of base material auxiliary laser deabsorption includes fixing nanozeolite molecular sieve doped with heteroatom in bodyframe structure, dipping mass spectrum stainless steel sample target plate of base material auxiliary laser deasorption in titanate coupler, placing dipped target plate in anhydrous ethyl acetate to let heteroatom nanozeolite crystal material suspension liquid be sufficiently dispersed then adding said liquid in sample-set hole of said sample target plate and baking said sample target plate.
Description
Technical field
The present invention relates to a kind of method of technical field of biological, specifically is the method for preparing functional target disk of mass spectrum sample de-absorbed by ground substance assistant laser that low abundance Phosphorylated Peptide/phosphorylating protein molecule is identified in a kind of direct enrichment.
Background technology
The protein phosphorylation process is the most general in the biosome, most important protein post-translational modification mode.This reversible process of the phosphorylation of protein and dephosphorylation has participated in the higher eukaryotic cell signal transduction, nearly all vital movement processes such as cell differentiation and cell growth, and with the major disease of many threat human healths, closely related as the generation development of tumour etc.The purpose of modern proteomics research is exactly to study whole expression of gene in translation and back modification level, thereby from the component law of the level research protein of an overall dynamics.All can have been constituted the phosphorylating protein group by the protein of phosphorylation in the cell.
Although effectively discern and identify low abundance phosphorylating protein molecule, can resolve the cell activities process better.But up to now, the analysis of the full spectrum of dynamic phosphorylation and extensive, low abundance phosphorylating protein molecule is identified on high flux ground, remains association area " bottleneck " problems anxious to be solved such as life science.Traditional
32P radioactive label method has directly, and the characteristics that sensitivity is high detect but can only can carry out radiolabeled cell to part, owing to there is radioactive pollution, have greatly limited its further application simultaneously.The Western blotting detection rule that depends on antibody is to utilize anti-phosphorylation antibody mediated immunity separation detection phosphorylating protein.But with respect to anti-serine/anti-threonine phosphorylation antibody, the specificity of only anti-tyrosine phosphorylation antibody is better, and is also commonly used at present.
The fast development of modern detecting such as substance assistant laser desorpted mass spectrum is for the structure evaluation of hanging down abundance phosphorylating protein molecule provides new opportunity to develop.In the process, the main phosphated peptide section by existing in the evaluating objects sample enzymolysis potpourri is to determine existing of corresponding phosphorylating protein molecule.But for the protein of numerous and complicated in the actual living things system is formed, because it is very limited that the protein molecule ratio of phosphorylation takes place, therefore, effective concentration and separation process, and, be a sensitive gordian technique approach identifying low abundance phosphorylating protein molecule with the combination between substance assistant laser desorpted mass spectrometer.
As everyone knows, fixing metal ions affinity chromatography (IMAC) is according to being incorporated into glucosan, the metallic ion in the flexible glue matrix stationary phase carriers such as agarose, and the characteristic of selective adsorption phosphate group, thus reach the purpose of enriching phosphated peptide section.
Through the prior art literature search is found, Jamie etc. are at authoritative academic journal-" U.S.'s analytical chemistry " (2006 in international analytical chemistry field, 78,1574) go up report: on the corresponding stainless steel sample of substance assistant laser desorpted mass spectrometer target disc surface, behind crosslinked modification polyacrylic acid-triacetic acid nitrous acid compound, by the fixing Fe (III) of interionic electrostatic attraction effect, thereby with Fe (III) is the affine avtive spot of metallic ion, the phosphated peptide section that may exist in the direct enrichment target sample system, the existence of corresponding low abundance phosphorylating protein molecule is identified in the substance assistant laser desorpted mass spectrum research of later use.But it is to be noted, still exist in the said process: corresponding substance assistant laser desorpted mass spectrometer sample target disc manufacturing process is loaded down with trivial details, and by interionic electric charge graviational interaction secure bond Fe (III), be difficult to avoid in the actual mechanical process, problems such as Fe (III) avtive spot loss, the directly Analysis and Identification of disturbing effect target phosphorylating protein molecule.
Summary of the invention
The present invention is directed to the deficiencies in the prior art, a kind of method for preparing functional target disk of mass spectrum sample de-absorbed by ground substance assistant laser is provided.The present invention utilizes suitable inorganic nano package technique, in specific point sample hole, the MALDI of corresponding stainless steel sample target disc surface, modify the molecular sieve of Nano zeolite crystalline material that fixedly is doped with heteroatoms avtive spots such as iron-titanium in the skeleton structure one by one equably.New medium assisted laser desorption mass spectrum (MALDI) the function sample target disc that corresponding preparation provides, can overcome at present direct enrichment effectively and identify the technological difficulties problem that exists in the low abundance phosphorylating protein molecular process, thereby, be high flux, low abundance phosphorylating protein molecule in the actual living things system of Analysis and Identification on a large scale, the technological approaches of the complete novelty with actual scientific meaning is provided.
The present invention is achieved by the following technical solutions, and concrete steps are as follows:
(1) prepare the fixing heteroatomic molecular sieve of Nano zeolite that mixes in the skeleton structure by hydrothermal synthesis method, corresponding heteroatoms nanosized zeolitic material, X ray polycrystalline diffraction analysis shows that characteristic diffraction peak shape is sharp-pointed, product crystallinity is good.Utilize deionized water rinsing repeatedly, to the nearly neutrality of respective sample aaerosol solution, oven dry, roasting, after impregnated in the ethyl acetate of no water treatment, it is stand-by that ultrasonic dispersion is prepared against;
In described (1), prepare the fixing heteroatomic molecular sieve of Nano zeolite that mixes in the skeleton structure, be specially by hydrothermal synthesis method:
Used silicon source is a white carbon; Used organic formwork agent is tetraethyl oxyammonia or tetrapropyl oxyammonia, under the titanium source participates in, adds iron chloride or gallium chloride, and hydrothermal crystallizing obtains the fixing heteroatomic molecular sieve of Nano zeolite that mixes in the skeleton structure under 413K.
(2) get the above-mentioned aaerosol solution of accurate volume, oven dry according to the nt wt net weight of gained molecular sieve of Nano zeolite material, determines that tentatively ratio is 0.1 gram nanosized zeolitic material/100 milliliter ethyl acetate between the initial solid-liquid in this material aaerosol solution;
Described oven dry, temperature are 100 ℃.
(3) the material aaerosol solution that obtains by ethyl acetate stepwise dilution above-mentioned (2) of back is to original 1/10,0000-1/10000 times;
(4) in ethanol-acetone organic solvent that the 1:1 volume ratio is formed, recycle the MALDI sample target disc that stainless steel is cleaned in the ultrasonic cleaner vibration, after utilize the 2mol/L diluted hydrochloric acid aqueous solution under 30 ℃ of condition of water bath heating, flood corresponding M ALDI sample target disc cmpletely, duration is 7-8 hour, MALDI sample target disc is washed repeatedly with secondary deionized water in the back, after thoroughly purging sample target disc surface moisture content with ear washing bulb, 100 ℃ of oven dry, be impregnated in the mixed solution of titanate coupling agent that volume ratio is 1:1000-5:1000 and anhydrous ethyl acetate 1 hour then, MALDI sample target disc is taken out in the back, aeration-drying under the room temperature, the low boiling ethyl acetate that may exist is removed in 80-100 ℃ of insulation with heating in baking oven.Thereby, evenly be coated with stain one deck titanate coupling agent on corresponding MALDI stainless steel sample target disc surface;
The titanate coupling agent of selected use comprises a kind of in positive four butyl esters of metatitanic acid, monoalkoxy fatty acid titanate esters, the monoalkoxy unsaturated fatty acid titanate esters.
(5) in the sample hose of miniature point gum machine, add the nano zeolite crystalline material aaerosol solution that above-mentioned (3) dilution obtains.Even the sample hose of nano zeolite crystalline material aaerosol solution is housed fully to be immersed in the miniature ultrasonic washer, answer the nano zeolite crystalline material by the abundant disperse phase of ultrasound wave, and remove the tiny bubble that may exist in the solution, under the state that keeps nano zeolite crystalline material aaerosol solution fully to disperse, join fast one by one in the point sample hole of handling the MALDI sample target disc that obtains in above-mentioned (4) then;
In described (5), the nano zeolite crystalline material aaerosol solution addition that dilution obtains is 1 milliliter; The volume of miniature ultrasonic washer is 100 milliliters, and the ultrasound wave frequency of operation is 40KHz, and the submergence temperature is 30 ℃.
(6) the MALDI sample target disc that above-mentioned (5) are obtained steadily places baking oven insulation back to take out, by the even titanate coupling agent of modifying in MALDI sample target disc surface, with the metal heteroatom interphase interactions such as iron-titanium of mixing and having in the heteroatoms molecular sieve of Nano zeolite crystalline material skeleton structure in the corresponding aaerosol solution, evenly modify the nano zeolite crystalline material that fixedly is doped with metal heteroatoms such as iron-titanium in the skeleton structure on MALDI sample target disc surface.
In described (6), baking oven is warming up to 80-100 ℃ in advance, and MALDI sample target disc is incubated 5 hours therein.
The MALDI sample target disc that the present invention makes, directly low abundance Phosphorylated Peptide/phosphorylating protein molecule in the target sample system is identified in enrichment.
The present invention is according to prior art, and make full use of the unique advantage that the heteroatoms molecular sieve of Nano zeolite crystalline material that fixedly is doped with metal active sites such as iron-titanium in the skeleton structure is had: outside surface enriches controlled; The duct is evenly abundant, good rigidly, and physical and chemical stability is strong; The easy modulation of macro morphology; Special natures such as easy derivatization.In the experiment, regulate and control key influence factors such as corresponding inorganic nano molecular sieve assembly material quantity, pattern, structure composition by microcosmic, utilize certain inorganic nano package technique, on MALDI stainless steel sample target disc surface, the uniformly fixing heteroatomic molecular sieve of Nano zeolite crystalline material such as doping iron-titanium of modifying in the suitable skeleton structure of thickness fixedly.The novel MALDI function sample target disc that corresponding preparation provides, it is simple and convenient to have manufacturing process, good reproducibility can effectively satisfy and use the needs that the low abundance phosphorylating protein molecule that may exist in the complicated actual sample system is identified in the direct enrichment of MALDI-MS instrument.
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The installation drawing that Fig. 1 adopts for the embodiment of the invention
Embodiment
Below in conjunction with accompanying drawing embodiments of the invention are elaborated: present embodiment is being to implement under the prerequisite with the technical solution of the present invention, provided detailed embodiment and concrete operating process, but protection scope of the present invention is not limited to following embodiment.
Example 1-3: in the building-up process of relevant nanometer zeolite molecular sieve, by modulation reactant FeCl
3The proportioning addition of solution, to obtain fixedly being doped with 0.1% respectively in the skeleton structure, 0.5%, 1.0%, 1.5%, 2.0%, 2.5%, 3.0%, 3.5%, 4.0%, the heteroatoms nano zeolite crystalline material (the crystalline size size is about 250-300nm) of 4.5% a series of different proportion ferrous metal elements, the back makes the synthetic heteroatoms nano zeolite crystalline material that obtains according to concrete technology path described in invention technical scheme (2)-(6), evenly modify in MALDI stainless steel sample target disc surface, to prepare the surperficial novel MALDI function sample target disc that evenly is modified with the molecular sieve of Nano zeolite crystalline material of doping different proportion ferrous metal atom in the skeleton structure.The novel MALDI function of this kind of later use sample target disc, directly enrichment identifies that low abundance, mass number in the β casein enzymolysis potpourri are the existence of 2061 Phosphorylated Peptide.From mass spectrometry results, can find, select to be doped with in the skeleton structure nanosized zeolitic material of 3.5% ferrous metal atom in the experiment, and the MALDI sample function target disc for preparing, it for mass number in the β casein enzymolysis potpourri 2061 phosphated peptide section, can be by effective enrichment, the background signal interference problem be improved significantly.
Example 4: in the building-up process of relevant nanometer zeolite molecular sieve, by modulation reactant GaCl
3The proportioning addition of solution, obtain fixedly being doped with 0.1% respectively in the skeleton structure, 0.5%, 1.0%, 1.5%, 2.0%, 2.5%, 3.0%, 3.5%, the heteroatoms nano zeolite crystalline material (the crystalline size size is about 250-500nm) of 4.0% a series of different proportion gallium elements, the back makes fixedly to be doped with the nano zeolite crystalline material of different proportion gallium atom as avtive spot in the skeleton structure according to concrete technology path described in invention technical scheme (2)-(6), evenly modifies in MALDI stainless steel sample target disc surface, prepare MALDI function sample target disc, directly low abundance in the β casein enzymolysis potpourri is identified in enrichment, mass number is 2061.062 Phosphorylated Peptide.Relevant experimental result shows, select the nano zeolite crystalline material of the fixing 3.5% gallium atom that mixes in the skeleton structure, and the MALDI sample function target disc for preparing can direct effectively enrichment be identified low abundance phosphated peptide section in the proteolysis potpourri.
Example 5: by source of iron in the modulation relevant nanometer zeolite molecular sieve synthesis reaction mixture, the titanium source, heteroatoms material proportion of composing such as gallium source, obtain in the skeleton structure simultaneously fixedly the iron of doping different proportion-titanium gallium-titanium thermometal heteroatoms as the nano zeolite crystalline material of avtive spot, according to concrete technology path described in invention technical scheme (2)-(6), the above-mentioned synthetic molecular sieve of Nano zeolite material that obtains is evenly modified in MALDI stainless steel sample target disc surface, prepare MALDI function sample target disc, directly low abundance in the β casein enzymolysis potpourri is also identified in enrichment simultaneously, mass number is 2061.062 Phosphorylated Peptide, and the existence of hanging down the abundance phosphated peptide section in the proteolysis potpourri of other actual sample system.
Example 6: in the building-up process of relevant nanometer zeolite molecular sieve, by the temperature that changes hydrothermal synthesis reaction, the time of reaction, and obtain a series of different crystal sizes, fixedly be doped with the iron, titanium atom of suitable proportion nano zeolite crystalline material in the skeleton structure as avtive spot.Relevant experimental result shows, is 413K when selecting the hydrothermal synthesis reaction temperature, and the reaction time is that the relevant nanometer zeolite molecular sieve crystal range of size that obtains is approximately 350-500nm under 72 hours the condition.Thereby, according to concrete technology path described in invention technical scheme (2)-(6), making the above-mentioned synthetic range of size that obtains is the molecular sieve of Nano zeolite material of 350-700nm, evenly modify in MALDI stainless steel sample target disc surface, prepare MALDI function sample target disc, the effect that low abundance Phosphorylated Peptide in the proteolysis potpourri is identified in its direct enrichment is satisfactory.
Example 7: according to the molecular sieve of Nano zeolite material of elements such as doping iron in the synthetic skeleton structure that obtains as avtive spot, utilize the formic acid-ammonium formate buffer solution of pH value in the 8.0-9.0 scope, dilution obtains the nano zeolite crystalline material aaerosol solution of a series of different solid ratios, join in the point sample hole of MALDI sample target disc, according to the detailed process described in the invention technical scheme, prepare a series of MALDI function sample target discs that are modified with different absolute magnitude nano zeolite crystalline materials.Experimental result shows, when making the solid-liquid ratio is the molecular sieve of Nano zeolite aaerosol solution of 0.1 gram nanosized zeolitic material/100 milliliter anhydrous ethyl acetate, be diluted to 1/10 by anhydrous ethyl acetate, in the time of 0000 times, the corresponding MALDI sample function target disc for preparing, observe under the polarizing microscope of 200 times of enlargement factors and find that the nano crystal particles of sample target disc finishing is uniform sequential, the effect that low abundance phosphated peptide section in the proteolysis potpourri is identified in its direct enrichment is satisfactory.
Example 8-9: by the water-bath heating arrangement of control miniature ultrasonic washer, the sample aaerosol solution that relevant nanometer zeolite molecular sieve material in the sample hose that is contained in micro-automatically dropping glue machine that is immersed in is fully wherein formed, respectively at 25 ℃, 30 ℃, 35 ℃, 40 ℃, 45 ℃, under 50 ℃ of a series of condition of different temperatures, join in the corresponding M ALDI point sample hole, with according to concrete technology path described in the invention technical scheme, prepare MALDI function sample target disc.Experimental result shows that 35 ℃ is that assurance relevant nanometer zeolite crystal material is evenly modified in the preference temperature on MALDI sample target disc surface.In the point sample hole of the corresponding MALDI sample target disc that obtains, be modified with thickness equably and suit, fixedly be doped with the heteroatomic nano zeolite crystalline material of iron-titanium in the skeleton structure simultaneously.
Example 9-10: according to concrete technology path described in invention technical scheme (1)-(6), from concrete Experimental equipment shown in Figure 1, make the sample hose of the commercial micro-automatically dropping glue machine of the relevant nanometer zeolite molecular sieve material aaerosol solution that is loaded with suitable concentration, in now being positioned over corresponding miniature ultrasonic cleaner water-bath, in the ultrasound wave frequency of operation is under the 40KHz condition, fully disperse corresponding nano zeolite crystal suspending liquid, satisfy with the nanocrystal aaerosol solution that guarantees to join in the MALDI function sample target disc point sample hole: the abundant precondition of disperseing and removing the tiny bubble that wherein may exist fully, successfully to prepare MALDI function sample target disc.
Example 11-12: in ethanol-acetone organic solvent that the 1:1 ratio is formed, recycle the MALDI sample target disc that stainless steel is cleaned in the ultrasonic cleaner vibration, after utilize the 2mol/L diluted hydrochloric acid aqueous solution under 30 ℃ of condition of water bath heating, flood corresponding M ALDI sample target disc cmpletely, duration is about 7-8 hour, take out MALDI sample target disc then, purge the moisture content that its surface may exist gently with ear washing bulb, MALDI sample target disc is washed repeatedly with secondary deionized water in the back, after thoroughly purging sample target disc surface moisture content with ear washing bulb, 100 ℃ of oven dry, be impregnated in titanate coupling agent then: 1h in the mixed solution of anhydrous ethyl acetate (volume ratio is 1:1000), MALDI sample target disc is taken out in the back, aeration-drying under the room temperature, the low boiling ethyl acetate that may exist is removed in 90 ℃ of insulations with heating in baking oven; Thereby, evenly be coated with stain one deck titanate coupling agent on corresponding MALDI stainless steel sample target disc surface.Process is described according to (5)-(6) step in the example (1) in the back, successfully prepares the MALDI function sample target disc that the surface evenly is modified with the heteroatoms nano crystal material;
Example 13: in the building-up process of relevant nanometer zeolite molecular sieve, selecting white carbon is the silicon source, and the tetraethyl oxyammonia is an organic formwork agent, under the titanium source participates in, add iron chloride/gallium chloride, hydrothermal crystallizing obtains the fixing heteroatomic molecular sieve of Nano zeolite that mixes in the skeleton structure under 413K; According to concrete technology path described in invention technical scheme (2)-(6), making the above-mentioned synthetic range of size that obtains is the molecular sieve of Nano zeolite material of 350-700nm, evenly modify in MALDI stainless steel sample target disc surface, prepare MALDI function sample target disc, the effect that low abundance Phosphorylated Peptide in the proteolysis potpourri is identified in its direct enrichment is satisfactory.
Example 14: in the building-up process of relevant nanometer zeolite molecular sieve, selecting white carbon is the silicon source, and the tetrapropyl oxyammonia is an organic formwork agent, under the titanium source participates in, add iron chloride/gallium chloride, hydrothermal crystallizing obtains the fixing heteroatomic molecular sieve of Nano zeolite that mixes in the skeleton structure under 413K; According to concrete technology path described in invention technical scheme (2)-(6), making the above-mentioned synthetic range of size that obtains is the molecular sieve of Nano zeolite material of 350-700nm, evenly modify in MALDI stainless steel sample target disc surface, prepare MALDI function sample target disc, the effect that low abundance Phosphorylated Peptide in the proteolysis potpourri is identified in its direct enrichment is satisfactory.
Example 15: select positive four butyl esters of metatitanic acid as titanate coupling agent, anticipate MALDI stainless steel sample target disc surface.According to concrete technology path described in invention technical scheme (2)-(6), make corresponding heteroatoms molecular sieve of Nano zeolite crystalline material, evenly modify in MALDI stainless steel sample target disc surface, prepare MALDI function sample target disc, the effect that low abundance Phosphorylated Peptide in the proteolysis potpourri is identified in its direct enrichment is satisfactory.
Example 16: select monoalkoxy fatty acid titanate esters as titanate coupling agent, anticipate MALDI stainless steel sample target disc surface.According to concrete technology path described in invention technical scheme (2)-(6), make corresponding heteroatoms molecular sieve of Nano zeolite crystalline material, evenly modify in MALDI stainless steel sample target disc surface, prepare MALDI function sample target disc, the effect that low abundance Phosphorylated Peptide in the proteolysis potpourri is identified in its direct enrichment is satisfactory.
Example 17: select monoalkoxy unsaturated fatty acid titanate esters as titanate coupling agent, anticipate MALDI stainless steel sample target disc surface.According to concrete technology path described in invention technical scheme (2)-(6), make corresponding heteroatoms molecular sieve of Nano zeolite crystalline material, evenly modify in MALDI stainless steel sample target disc surface, prepare MALDI function sample target disc, the effect that low abundance Phosphorylated Peptide in the proteolysis potpourri is identified in its direct enrichment is satisfactory.
Example 18: according to concrete technology path described in invention technical scheme (1)-(6), after making ordinary laboratory be warming up to 80 ℃ in advance with baking oven, add relevant nanometer crystal aaerosol solution in the point sample hole with MALDI sample target disc one by one, back smooth transfer places baking oven, continue evenly to modify the MALDI function sample target disc of heteroatoms nano crystal material successfully to obtain the surface 80 ℃ of insulation taking-ups after 5 hours down.
Example 19-20: according to the MALDI function sample target disc for preparing, point adds proteolysis hybrid peptide section, selecting DHA is the matrix micromolecule that uses in the MALDI-TOF-MS process, utilize acetonitrile-acetic acid mixed solution, the non-phosphorylating peptide section of sample target disc surface non-specific binding is removed in drip washing, identify the Phosphorylated Peptide in the proteolysis products such as the high phosphoprotein of ovum gallinaceum, β casein with direct enrichment, and utilize MALDI-TOF-MS to identify the existence of phosphorylation site in the respective egg white matter molecular structure.
Claims (10)
1, a kind of method for preparing functional target disk of mass spectrum sample de-absorbed by ground substance assistant laser is characterized in that, concrete steps are as follows:
(1) prepares the fixing heteroatomic molecular sieve of Nano zeolite that mixes in the skeleton structure by hydrothermal synthesis method, utilize deionized water rinsing repeatedly, to the nearly neutrality of respective sample aaerosol solution, oven dry, roasting, after impregnated in the ethyl acetate of no water treatment, ultrasonic dispersion is in order to stand-by;
(2) get above-mentioned aaerosol solution oven dry,, determine that ratio is 0.1 gram nanosized zeolitic material/100 milliliter ethyl acetate between the initial solid-liquid in this material aaerosol solution according to the nt wt net weight of gained molecular sieve of Nano zeolite material;
(3) utilize material aaerosol solution that ethyl acetate dilution above-mentioned (2) obtains to original 1/100000-1/10000 doubly;
(4) in ethanol-acetone organic solvent that the 1:1 volume ratio is formed, recycle the ultrasonic cleaner vibration and clean substance assistant laser desorpted mass spectrum sample target disc, after utilize aqueous hydrochloric acid solution under water bath condition, to flood substance assistant laser desorpted mass spectrum sample target disc, wash repeatedly with secondary deionized water after taking out substance assistant laser desorpted mass spectrum sample target disc, after purging surperficial moisture content with ear washing bulb, oven dry, after impregnated in the mixed solution of titanate coupling agent and anhydrous ethyl acetate, substance assistant laser desorpted mass spectrum sample target disc is taken out in the back, aeration-drying under the room temperature, the low boiling ethyl acetate that may exist is removed in insulation in baking oven;
(5) in the sample hose of miniature point gum machine, add the nano zeolite crystalline material aaerosol solution that above-mentioned (3) dilution obtains, sample hose fully is immersed in the miniature ultrasonic washer, by ultrasonic dispersing nano zeolite crystalline material, and remove tiny bubble in the solution, under the state that keeps nano zeolite crystalline material aaerosol solution fully to disperse, join fast one by one in the point sample hole of substance assistant laser desorpted mass spectrum sample target disc then;
(6) the substance assistant laser desorpted mass spectrum sample target disc that above-mentioned (5) are obtained steadily places baking oven insulation back to take out, by the even titanate coupling agent of modifying in substance assistant laser desorpted mass spectrum sample target disc surface, with the metal heteroatom interphase interaction of mixing and having in the heteroatoms molecular sieve of Nano zeolite crystalline material skeleton structure in the corresponding aaerosol solution, evenly modify the nano zeolite crystalline material that is doped with metal heteroatom in the skeleton structure on substance assistant laser desorpted mass spectrum sample target disc surface.
2, the preparation method of a kind of functional target disk of mass spectrum sample de-absorbed by ground substance assistant laser according to claim 1, it is characterized in that, in described (1), prepare the fixing heteroatomic molecular sieve of Nano zeolite that mixes in the skeleton structure, be specially by hydrothermal synthesis method:
Used silicon source is a white carbon; Used organic formwork agent is tetraethyl oxyammonia or tetrapropyl oxyammonia, under the titanium source participates in, adds iron chloride or gallium chloride, and hydrothermal crystallizing obtains the fixing heteroatomic molecular sieve of Nano zeolite that mixes in the skeleton structure under 413K.
3, the preparation method of a kind of functional target disk of mass spectrum sample de-absorbed by ground substance assistant laser according to claim 1 is characterized in that, in described (1), bake out temperature is 100 ℃.
4, the preparation method of a kind of functional target disk of mass spectrum sample de-absorbed by ground substance assistant laser according to claim 1 is characterized in that, in described (1), roasting is meant: at 550 ℃ of roasting 3-4h.
5, a kind of method for preparing functional target disk of mass spectrum sample de-absorbed by ground substance assistant laser according to claim 1, it is characterized in that, in described (4), utilize aqueous hydrochloric acid solution under water bath condition, to flood substance assistant laser desorpted mass spectrum sample target disc, wherein: the concentration of aqueous hydrochloric acid solution is 2mol/L, bath temperature is 30 ℃, and the dipping duration is 7-8 hour.
6, a kind of method for preparing functional target disk of mass spectrum sample de-absorbed by ground substance assistant laser according to claim 1, it is characterized in that, in described (4), impregnated in the mixed solution of titanate coupling agent and anhydrous ethyl acetate, wherein: the volume ratio of titanate coupling agent and anhydrous ethyl acetate is 1:1000-5:1000 in the mixed solution of titanate coupling agent and anhydrous ethyl acetate; Dip time is 1 hour.
7, according to claim 1 or 6 described a kind of method for preparing functional target disk of mass spectrum sample de-absorbed by ground substance assistant laser, it is characterized in that the titanate coupling agent of selected use comprises a kind of in positive four butyl esters of metatitanic acid, monoalkoxy fatty acid titanate esters, the monoalkoxy unsaturated fatty acid titanate esters.
8, a kind of method for preparing functional target disk of mass spectrum sample de-absorbed by ground substance assistant laser according to claim 1 is characterized in that, in described (4), bake out temperature is 100 ℃, and holding temperature is 80-100 ℃.
9, the preparation method of a kind of functional target disk of mass spectrum sample de-absorbed by ground substance assistant laser according to claim 1 is characterized in that, in described (5), the nano zeolite crystalline material aaerosol solution addition that dilution obtains is 1 milliliter; The volume of miniature ultrasonic washer is 100 milliliters, and the ultrasound wave frequency of operation is 40KHz, and the submergence temperature is 30 ℃.
10, the preparation method of a kind of functional target disk of mass spectrum sample de-absorbed by ground substance assistant laser according to claim 1, it is characterized in that, in described (6), baking oven is warming up to 80-100 ℃ in advance, and substance assistant laser desorpted mass spectrum sample target disc is incubated 5 hours therein.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102798658A (en) * | 2012-07-20 | 2012-11-28 | 浙江大学 | Method for analyzing low and medium-abundance low molecular weight protein in serum |
| WO2016116012A1 (en) * | 2015-01-21 | 2016-07-28 | 华中师范大学 | Mass correction kit and correction method for low-mass area of high-resolution mass spectrometer in negative ion mode |
Families Citing this family (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102899724B (en) * | 2011-09-15 | 2016-05-11 | 江苏有能光电科技有限公司 | A kind of method of eliminating bubble in sapphire crystal growth process |
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-
2007
- 2007-01-25 CN CNB2007100367896A patent/CN100504378C/en not_active Expired - Fee Related
Non-Patent Citations (2)
| Title |
|---|
| 运用生物质谱分析磷酸化多肽合成中的主产物. 寻晓红等.东莞理工学院学报，第6卷第3期. 2006 |
| 运用生物质谱分析磷酸化多肽合成中的主产物. 寻晓红等.东莞理工学院学报，第6卷第3期. 2006 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102798658A (en) * | 2012-07-20 | 2012-11-28 | 浙江大学 | Method for analyzing low and medium-abundance low molecular weight protein in serum |
| WO2016116012A1 (en) * | 2015-01-21 | 2016-07-28 | 华中师范大学 | Mass correction kit and correction method for low-mass area of high-resolution mass spectrometer in negative ion mode |
| US10181395B2 (en) | 2015-01-21 | 2019-01-15 | Central China Normal University | Mass calibration kit and calibration method for low-mass region of high-resolution mass spectrometer in negative ion mode |
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